Enrichment of Umbilical Cord Blood Mononuclears with Hemopoietic Precursors in Co-Culture with Mesenchymal Stromal Cells from Human Adipose Tissue

Maslova, Elena V.; Andreeva, E. R.; Андрианова, И. В.; Бобылева, П. И.; Romanov, Yu. A.; Kabaeva, N. V.; Balashova, Elena E.; Ryaskina, S. S.; Dugina, T. N.; Буравкова, Л. Б.

doi:10.1007/s10517-014-2400-9

Cited by 12 publications

(6 citation statements)

References 21 publications

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“…Cord blood MNCs are a heterogeneous population of blood-borne cells with the vast majority of mature blood-borne elements and a small subpopulation of cbHSPC progenitors of different commitment. In this paper, we used the “physiological” approach to enrich cbHSPC fractions, as described earlier [ 28 ]. The total cbMNCs were co-cultured with ASC feeder for 3 days, after which all of the non-attached cells were removed.…”

Section: Discussionmentioning

confidence: 99%

“…To enrich the population of cbHSPCs we used the experimental approach described by us previously [ 28 ]. Briefly, confluent (70–80%) ASC layers were pre-formed in 35 mm Petri dishes at 20% and 5% O 2 .…”

Section: Methodsmentioning

confidence: 99%

“…This may lead to artificial depletion of the heterogeneous HSPC population, as this antigen is expressed on only some subsets of HSPCs [ 27 ]. To enrich selection of the HSPC population from cord blood, we applied a functional approach at the expense of HSPC adhesion to the ASC layer and further expansion of the adhered HSPCs and their progeny [ 28 ]. In this study, we evaluated for the first time the potency of ASCs to support viability and self-renewal/commitment of expanded HSPCs (progeny of cells which were adhered to unmanipulated cbMNCs) at standard (20%) and tissue-related (5%—hypoxia) O 2 .…”

Section: Introductionmentioning

confidence: 99%

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Human Adipose-Tissue Derived Stromal Cells in Combination with Hypoxia Effectively Support Ex Vivo Expansion of Cord Blood Haematopoietic Progenitors

et al. 2015

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The optimisation of haematopoietic stem and progenitor cell expansion is on demand in modern cell therapy. In this work, haematopoietic stem/progenitor cells (HSPCs) have been selected from unmanipulated cord blood mononuclear cells (cbMNCs) due to adhesion to human adipose-tissue derived stromal cells (ASCs) under standard (20%) and tissue-related (5%) oxygen. ASCs efficiently maintained viability and supported further HSPC expansion at 20% and 5% O2. During co-culture with ASCs, a new floating population of differently committed HSPCs (HSPCs-1) grew. This suspension was enriched with СD34+ cells up to 6 (20% O2) and 8 (5% O2) times. Functional analysis of HSPCs-1 revealed cobble-stone area forming cells (CAFCs) and lineage-restricted colony-forming cells (CFCs). The number of CFCs was 1.6 times higher at tissue-related O2, than in standard cultivation (20% O2). This increase was related to a rise in the number of multipotent precursors - BFU-E, CFU-GEMM and CFU-GM. These changes were at least partly ensured by the increased concentration of MCP-1 and IL-8 at 5% O2. In summary, our data demonstrated that human ASCs enables the selection of functionally active HSPCs from unfractionated cbMNCs, the further expansion of which without exogenous cytokines provides enrichment with CD34+ cells. ASCs efficiently support the viability and proliferation of cord blood haematopoietic progenitors of different commitment at standard and tissue-related O2 levels at the expense of direct and paracrine cell-to-cell interactions.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Human Adipose-Tissue Derived Stromal Cells in Combination with Hypoxia Effectively Support Ex Vivo Expansion of Cord Blood Haematopoietic Progenitors

et al. 2015

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“…Isolation of CBMNCs was performed as reported before [21]. In brief, 35ml of fresh cord blood was collected from a healthy woman with an uncomplicated delivery.…”

Section: Cord Blood Mononuclear Cells (Cbmncs) Isolationmentioning

confidence: 99%

Co-transplantation of Hypoxia Pretreated Human Adipose Derived Mesenchymal Stem Cells and Cord Blood Mononuclear Cells to Treat Rats with Acute Myocardial Infarction

Xiang¹,

Xiang²,

Zhang³

et al. 2020

Preprint

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BackgroundHuman adipose derived mesenchymal stem cells (ASCs) are ideal candidates for the treatment of acute myocardial infarction (AMI), due to their favorable availability and regenerative potential. However, in vivo studies showed that ASCs are not resilient at the infarcted area, for a shortage of blood and oxygen supply. Hypoxic pretreatment was proven to be an effective way to enhance cell survival in ischemic atmosphere. Moreover, co-transplantation of stem cells was another promising strategy to improve cardiac function after transplantation. So, we hypothesized that hypoxic pretreated ASCs combined with proangiogenic cord blood mononuclear cells (CBMNCs) would promote treatment efficacy after co-transplantation.MethodsASCs extracted from male volunteer were preconditioned in hypoxic condition (HP-ASC) for 24h, and total RNA were extracted after that. Gene expressions were compared between HP-ASC and ASC. Then, we transplanted stem cells to female Wistar rats which divided into different groups: (1) HP-ASCs group (n=10, 1x106ASCs); (2) HP-ASCs + CBMNCs group (n=10, 0.5×106 ASCs+0.5×106 CBMNCs); (3) CBMNCs group (n=10, 1×106 ASCs); (4) Control group (n=10, 40μL PBS); (5) Sham group (n=10). Echocardiogram was performed before (0d) and after (30d) after cell transplantation. Hearts were harvested at 30d to analyze the infarct size, myocardium apoptosis, stem cells viability and angiogenesis. ResultsIn vitro study showed that HP-ASCs had a wide range of paracrine function, with the incretion growth factors and their receptors, which would support the cell survivals. In addition, HP-ASCs also gained potentials in hypoxic adaptation (increased expression of HO-1 and SDF-1), as well as homing and immigrating abilities (CXCR4, ICAM-1 and ICAM-2). In vivo studies showed that, 30 days after transplantation in AMI rats, the HP-ASCs group had a better improvement in cardiac function; reduction of the infarct size; and decrease of ASCs death than the other groups (HP-ASCs > HP-ASCs + CBMNCs ≧ CBMNCs > PBS) (p<0.05). However, the combined group of HP-ASCs and CBMNCs had more significant angiogenesis than the other groups (HP-ASCs + CBMNCs > CBMNCs > HP-ASCs > PBS) (p <0.05).ConclusionsHP-ASCs alone had a greater potential in improving cardiac function in AMI rats. However, the combination of HP-ASCs and CBMNCs had a better result in angiogenesis.

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“…MSCs from different sources differ in the expression of some markers, their ability to proliferate and differentiate, but in general their characteristics are similar [ 69 - 72 ]. MSCs from the stromal-vascular fraction of human adipose tissue have been shown to support hematopoiesis in vitro [ 53 , 73 ]. Therefore, they are a good alternative to bone marrow MSCs and represent an easily accessible source of feeder cells for the expansion of UCB HSPCs for widespread clinical use [ 74 ].…”

Section: Modeling a Specific Microenvironment For Ex Vivo Expansion Of Ucb Hspcsmentioning

confidence: 99%

Ex Vivo Expansion of Hematopoietic Stem and Progenitor Cells from Umbilical Cord Blood

et al. 2016

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Transplantation of umbilical cord blood cells is currently widely used in modern cell therapy. However, the limited number of hematopoietic stem and progenitor cells (HSPCs) and prolonged time of recovery after the transplantation are significant limitations in the use of cord blood. Ex vivo expansion with various cytokine combinations is one of the most common approaches for increasing the number of HSPCs from one cord blood unit. In addition, there are protocols that enable ex vivo amplification of cord blood cells based on native hematopoietic microenvironmental cues, including stromal components and the tissue-relevant oxygen level. The newest techniques for ex vivo expansion of HSPCs are based on data from the elucidation of the molecular mechanisms governing the hematopoietic niche function. Application of these methods has provided an improvement of several important clinical outcomes. Alternative methods of cord blood transplantation enhancement based on optimization of HPSC homing and engraftment in patient tissues have also been successful. The goal of the present review is to analyze recent methodological approaches to cord blood HSPC ex vivo amplification.

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Enrichment of Umbilical Cord Blood Mononuclears with Hemopoietic Precursors in Co-Culture with Mesenchymal Stromal Cells from Human Adipose Tissue

Cited by 12 publications

References 21 publications

Human Adipose-Tissue Derived Stromal Cells in Combination with Hypoxia Effectively Support Ex Vivo Expansion of Cord Blood Haematopoietic Progenitors

Human Adipose-Tissue Derived Stromal Cells in Combination with Hypoxia Effectively Support Ex Vivo Expansion of Cord Blood Haematopoietic Progenitors

Co-transplantation of Hypoxia Pretreated Human Adipose Derived Mesenchymal Stem Cells and Cord Blood Mononuclear Cells to Treat Rats with Acute Myocardial Infarction

Ex Vivo Expansion of Hematopoietic Stem and Progenitor Cells from Umbilical Cord Blood

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